Discovery of a potent, Kv7.3-selective potassium channel opener from a Polynesian traditional botanical anticonvulsant

The voltage-gated potassium (Kv) channel formed by Kv7.2/3 (KCNQ2/3) heteromers is a major target for anticonvulsant drug development. Here, we screened 1,444 extracts primarily from plants collected in California and the US Virgin Islands, for their ability to activate Kv7.2/3 but not inhibit Kv1.3, to select against tannic acid being the active component. We validated the 7 strongest hits, identified Thespesia populnea (miro, milo, portia tree) as the most promising, then discovered its primary active metabolite to be gentisic acid (GA). GA highly potently activated Kv7.2/3 (EC50, 2.8 nM). GA is, uniquely, 100% selective for Kv7.3 versus other Kv7 homomers; it requires S5 residue Kv7.3-W265 for Kv7.2/3 activation, and it ameliorates pentylenetetrazole-induced seizures in mice. Also an active aspirin metabolite, GA provides a molecular rationale for the use of T. populnea as an anticonvulsant in Polynesian indigenous medicine and presents novel pharmacological prospects for potent, iso..., Collection of plant samplesWe collected, between 2019 and 2022, aerial parts of plants under permit from Mojave National Preserve (study # MOJA-00321), Yosemite National Park (study # YOSE-00839), Santa Monica Mountains National Recreation Area (study # SAMO-00192), Muir Woods National Monument (study # MUWO-00035), Santa Cruz Island and Santa Rosa Island (study # CHIS-0023), and Boyd Deep Canyon (Indian Wells, CA) (permit through Philip L. Boyd Deep Canyon. Desert Research 376 Center, University of California, Riverside, CA) in California, US. In USVI, we collected from Virgin Islands National Park, St. John (study # VIIS-20001), Salt River Bay National Historical Park and Ecological Preserve, St. Croix (study # SARI-00056) and Buck Island Reef National Monument, St. Croix (study # BUIS-00103). Plant samples were collected in a manner designed to not kill the remaining plant, sealed in Ziploc bags (SC Johnson, Racine, WI, US), kept cold, and frozen as soon as possible. Other plant extr..., , ## Source Data for manuscript: Discovery of a potent, Kv7.3-selective potassium channel opener from a Polynesian traditional botanical anticonvulsant. The datasets included are the original Excel files used to generate each panel for figures 1-8 in this manuscript. The title of each Excel file is labeled to directly correspond to the figure in the manuscript: Figure Number & panel > Channel Investigated > Condition > Parameter Measured --- The types of files included within this repository are as follows: > Plant Screen > This is a database providing the full details of all 1444 plant extracts screened for activity against Kv7.2/7.3 and Kv1.3 as summarized by the dot-plot in Figure 1A and Figure 1B. The database contains columns providing identifying information for each plant where appropriate. The information provided in each column is as follows: Plant ID: Unique Identifier of the plant sample. The code corresponds to the region the sample was obtained and th...由Kv7.2/3（KCNQ2/3）异聚体构成的电压门控钾（voltage-gated potassium, Kv）通道是抗惊厥药物研发的核心靶点。本研究针对1444种提取物（主要采自美国加利福尼亚州与美属维尔京群岛的植物）开展筛选，以筛选可激活Kv7.2/3但不抑制Kv1.3的活性组分，从而排除单宁酸作为有效成分的可能性。我们验证了7个活性最强的命中化合物，确认桐棉（Thespesia populnea，又称miro、milo、波多树）为最具潜力的候选物，随后发现其主要活性代谢产物为龙胆酸（gentisic acid, GA）。GA对Kv7.2/3具有极强的激活活性（半数有效浓度EC₅₀=2.8 nM）。值得注意的是，GA相较于其他Kv7同源多聚体，对Kv7.3具有100%的选择性；其激活Kv7.2/3需要Kv7.3的S5残基W265参与，且可缓解戊四氮诱导的小鼠癫痫发作。作为阿司匹林的活性代谢产物之一，GA为波利尼西亚传统医学中使用桐棉作为抗惊厥剂提供了分子层面的理论依据，并为强效、亚型选择性…… 植物样本采集 2019年至2022年间，我们通过官方许可，于美国加利福尼亚州的莫哈韦国家保护区（研究编号MOJA-00321）、优胜美地国家公园（研究编号YOSE-00839）、圣塔莫尼卡山脉国家游乐区（研究编号SAMO-00192）、缪尔伍兹国家纪念碑（研究编号MUWO-00035）、圣克鲁斯岛与圣罗莎岛（研究编号CHIS-0023）以及加利福尼亚州印第安韦尔斯的博伊德深峡谷（由加州大学河滨分校菲利普·L·博伊德深峡谷沙漠研究中心许可，编号376）采集植物地上部分。在美属维尔京群岛，我们于维尔京群岛国家公园圣约翰岛（研究编号VIIS-20001）、盐河湾国家历史公园与生态保护区圣克罗伊岛（研究编号SARI-00056）以及巴克岛礁国家纪念碑圣克罗伊岛（研究编号BUIS-00103）开展样本采集。植物样本采集以不损伤剩余植株为原则，将样本密封于Ziploc保鲜袋（庄臣公司，SC Johnson，美国威斯康星州雷辛市）中，低温保存并尽快冷冻。其余植物提取物相关内容…… 本论文源数据：《Discovery of a potent, Kv7.3-selective potassium channel opener from a Polynesian traditional botanical anticonvulsant》（《从波利尼西亚传统植物抗惊厥剂中发现强效、Kv7.3选择性钾通道开放剂》） 本数据集包含用于生成本论文图1至图8各子图的原始Excel文件，每个Excel文件的命名规则均直接对应论文中的图表：`图编号&子图>研究通道>实验条件>检测参数`。 本仓库包含的文件类型如下： > 植物筛选数据库 > 该数据库完整收录了1444种植物提取物针对Kv7.2/7.3与Kv1.3的活性筛选详情，对应图1A与图1B的散点图汇总结果。数据库各列包含对应植物的必要识别信息，各列信息说明如下： > 植物ID：植物样本的唯一标识符，其编码对应样本采集区域及……